Abstract
Some mRNAs localize to specific regions within eukaryotic cells to express their functions. The movement and localization of mRNA molecules provides valuable information about how they concentrate to particular regions. Recent technical advances in optical microscopy and image analysis algorithms enable real-time tracking of single mRNA molecules in living cells. This chapter presents the methods to visualize and track single β-actin mRNA molecules that localize at the leading edge of chicken embryo fibroblasts. Furthermore, this chapter presents an analysis approach for single-molecule tracking data to extract quantitative information about the microenvironments of the mRNA molecules.
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Acknowledgments
We thank Yasuhiro Sasuga, Kayoko Terada (Tokyo Metropolitan Institute of Medical Science) and Yoshie Harada (Kyoto University) for constructing the tandemly repeated MS2 tag sequences. This research was partly supported by a Grant-in-Aid for Scientific Research on Innovative Areas (21121004) and by the Japan Society for the Promotion of Science (JSPS) through its “Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST Program)” to T.F.
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Yamagishi, M., Shirasaki, Y., Funatsu, T. (2013). Single-Molecule Tracking of mRNA in Living Cells. In: Sousa, A., Kruhlak, M. (eds) Nanoimaging. Methods in Molecular Biology, vol 950. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-137-0_10
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DOI: https://doi.org/10.1007/978-1-62703-137-0_10
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